Esters of erythromycin oxime

ABSTRACT

Covers stable oxime esters of erythromycin oximes, which are active antibiotic agents.

Umted States Patent [191 [111 3,869,444

Freiherg Mar. 4, 1975 ESTERS OF ERYTHROMYCIN OXIME 75 I t 1 L Al F W k a I Primary Examiner-Johnnie R. Brown nvefn or esle an rel f au egan Assistant E.\aminerCary B. Owems Asslgneer Abpott fi O leS, North Attorney, Agent, or Firm-Robert IL. Niblack; Vincent Chlcago, III. A. Mallare; James L. Bailey {22] Filed: Oct. 10, 1972 [21] Appl. No.: 296,431

[57] ABSTRACT [52] U.S. Cl. 260/210 E, 424/180 [51] Int. Cl. C07c 47/18 C t bl f th 58 Field of Search 260/210 E 9 5 a 9 O 0mm,

whtch are HCHVB anttblotlc agents.

[56] References Cited UNITED STATES PATENTS 14 Claims, N0 Drawings 3,574,185 4/1971 Tamburasw et al 260/210 E ESTERS OF ERYTHROMYCIN OXIME cin oxime. More particularly, it relates to oxime esters 5 of erythromycin A and B oxime that are stable to solvolysis which have the formula cu, (cladinose) (ez 'ychrono lide) where R is hydrogen or hydroxy, andR is a bulky alkyl group or a substituted aryl group, said substituted aryl group having the structural formula functional groups which modify and improve the thera peutic effect of the parent antibiotic. It has now been discovered thatthe erythromycin derivatives of this invention are especially stable to solvolysis, and furthermore, that such stability increases asthe R group defined above becomes increasingly sterically hindered.

The compounds of the invention are preparedby the reaction of an acyl halide witherythromycinoxime. The reaction may be carried out in an inert solvent,

-such being definedas one which does not react substantially with reactant orproduct.

in general, the compounds of this invention are prepared by reacting an acyl halide of the formula RCOX where R is as hereinbefore defined, and where X is halide, preferably chloride, with erythromycin oxime. This latter compound. is disclosed in US. Pat. No. 3,478,014. The reaction is best carried out by mixing the reactants at a temperatureranging from about 20 to 30 C. for a periodof about 2 to. 12 hours, at which time the reactionis considered substantially complete. The reaction isbestcarried out in an inert solvent as noted above such as dichloromethaneand with a proton aceptor'such as triethylamine.

The following examples further illustrate compounds prepared according to this method. The numbers in parentheses following the chemical name identify the compounds by such'specific number, which may be alluded to in succeeding examples merely to identify said compounds by'number.

EXAMPLE I 9-(0)-Trimethylacetylerythromycin B Oxime (l To astirred solution of 2.00 g. (2.73 mmol.) of erythromycin B oximein 50 ml. of dry acetone was added 1.00 g. (l 1.9 mmol.) of NaHCO followed by 0.365 g. (3.02 mmol.) of trimethylacetyl chloride. The mixture was stirred for 6 hours at 25 C. andthen was poured into350 ml. of water containing 30' ml. of concentrated NH OH. The product was extracted withfour 30 ml. portions of CHCl The combined CHCl layer was dried over anhyd. Na SO and evaporated to leave 2.05 g. of crude product. The product was crystallized from a mixture of 20 ml. of acetone and 5 ml. of water to give 1.24 g. of ester 1: mp l33135.

The reaction scheme for the above is as follows:

3 CH3+CH3 o 0 CH3 NaHCO3 acefone 3 EXAMPLE u To a stirred solution of 7.00 g. (9.35 mmol.) of erythromycin A oxime and 2.84 g. (28.0 mmol.) of triethylamine in 150 ml. of CH Cl at 25 C. was added 1.83 (10.2 mmol.) of 2,4,6-trimethylbenzoyl chloride. The mixture was stirred briefly and then allowed to stand at 25 C. for 4 hours.

The mixture was transferred to a separatory funnel and washed with 80 ml. of water containing 7 ml. of concentrated NH OH. The CH C1 layer was dried over anhyd. Na SO and concentrated. The residue was crystallized from a mixture of 70 ml. of chloroform and 70 ml. of hexane to give 4.00 g. of the ester 2: mp. 132135.

Anal. calcd. for c um o 895151):

c. 63.06; H. 8.78; 3.|3; 0, 25.0 Found: C; 62.89; H, 8.91;

EXAMPLE Ill 9-(0)-(2,4,6-Trimethyl)benzoylerythromycin B Oxime In a manner similar to Example 11, 1.00 g. of erythromycin B oxime was reacted with 2,4,6- trimethylbenzoyl chloride. The crude product was crystallized from 10 ml. of acetone and 5.5 m1. of water to give 0.55 g. of 3: mp. l2 6129.

Anal. calcd. for C H- N O (879.199):

C, 64.21; H, 8.95; N, 3.19; O, 23.66 Found: C, 64.13; H, 8.96; N, 3.20; 0, 23.67.

The reaction scheme for Examples 11 and 111 is as follows:

4 EXAMPLE IV 9-(0)-(2,6 Dimethyl-4-carbomethoxy) benzoylerythromycin A Oxime (4) Anal. calcd. for cqgH'mNgo g (939.160):

C, 61.39; H, 8.37; N, 2.9 Found: C, 61.25; H, 8.54; N. 2.9

EXAMPLE V 9-(0)-(2,6-Dimethyl-4- carbomethoxy)benzoylerythromycin B Oxime (5) In a manner similar to Example 11 2.00 g. of erythro- 20 mycin B oxime was reacted with (2,6-dimethyl-4- carbomethoxy)benzoyl chloride. The crude product was crystallized from a mixture of 20 m1. of ethanol and 18 ml. of water to give 1.27 g. of 5: mp. l32l36.

Anal. calcd. for c u mo (923.161):

C, 62.45; H. 8 Found: C, 62.31; H,

The reaction scheme for Examples 1V and V is as follows:

CO2CH3 EXAMPLE V1 50 (2,6-Dimethyl-4-benzyloxycarbonyl)benzoyl Chloride A CH Cl solution of the acid chloride 6 was prepared in the following manner: To 100 ml. of CH C1 55 was added 0.67 ml. of (6.7 mmol.) of benzyl alcohol and 3.04ml. (22.2Immol.) of triethylamine. The mixture was stirredand'cooledto C. in. an ice-bath. Then, 1.09-ml. (6.10 mmol.) of 2*,6-dimethylterephthaloyl chloridewas added and. after minutes the icebath was removed. The mixture was allowed. to stand at 25 C. for 2hours. A sample removed at that time for infrared analysis showed bands at 1795 cm. (acid chloride) and at 1729 cm. (ester). Such solutions were stable for several hours, but were normally used immediately.

EXAMPLE VII 9-(0)-(2,6-Dimethyl-4-benzyloxycarbonyl) b'enzoylerythromycin B Oxime (7) To the CH CI solution of 6 (ExampleVI) was added 4.00 g. (5.55 mmol.) of erythromycin B oxime. The mixture was stirred briefly to dissolve the oxime and. then was allowed to stand at 25 C for 4 hours. After transferring to a separatory funnel, the mixture was washed with 50 ml. of water containing 3 ml. of concentrated NI-I OI-I; The solvent was dried over Na SO and evaporated to give 5.44 g. of crude 7.

Purification of the. crude product was accomplished by chromatography. A columnof 100 g. of Florisil was prepared in benzene and conditioned by eluting with 1.0 liter of benzene: 0.3 percent triethylamine; The sample was placed on thecolumnin ml. of that solvent andthe column was eluted according to the following schedule, 125 ml. fractions being collected: frac. 1-4, (pH/0.3% Et N; frac. 4-8, H/l% MeOH/0.- 3%Et N; frac. 9-12, H/2%/MeOH/0.3%Et N; frac. l3l6, H/4%MeOH/0.3%Et l l. Fractions 3-l4 were shown by tlc. to be pure and were combined to give 4.44 g. of a glass. A 2.56 g. sample was crystallized from 15 ml. of acetone and 6 ml. of water to give 1.96 g. of 7: mp. ll8-l22.

Anal. calcd. for C H N O (999.259):

C, 64.91; H, 8.27; N, 2.80; 0, 24.02. Found: C, 65.22; H, 8.44; N, 2.84; 0, 23.70.

EXAMPLE VIIII [2,6-Dimethyl-4-( N-n-butylcarbamyl) Ibcnzoyl Chloride (8) A CH CI solution of the acid chloride 8 was prepared in the following manner: To 150 ml. of (H Cl was added 0835' ml. (8. 1 8 mmol.) of n-butylamine and 4.58 ml. (32.8 mmol.) of triethylamine. The mixture was stirred and'cooled. to 0 C. inan ice-bath. Then", 1.48 ml. (8.18 mmol.)of 2,6-dimethylterephthaloyl chloride was added and after 20 minutes asample was removed. for infrared analysiswhich showed bands at 179-5 cm. (acid chloride) andat 1668 cm? (amide). This solution was used directly without purification.

EXAMPLEIX 9-(0)-[2,6-Dimethyl-4-(N-N- butylcarbamyl)]benzoylerythromycin B Oxime (9) To the CH CIQ solution of 8 (Example VIII) at 0 C. was added 6.00 g. (8.18 mmol.) of erythromycin B oxime. The mixture was stirred briefly to dissolve the oxime and then was allowed to stand at 25 C. for 4 hours. The solution was then washedw-ith ml. of watercontainingS ml. of concentratedNH OI-I. The

solvent was dried. over anhyd. Na sOgandevaporated.

to give 7.46 g. of crude product.

Purification of the crude product was accomplished by chromatography. A column of- 140 g. of silica gel was prepared in CH CI and made" basic with 1.5 liter of CH CI 1% Et N. Theisampleof crude 9 (7146' g.) was placed on the column in 20 ml. of 'CH Cl l %Et N and elution was begun with that: solvent. Fractions of 125 ml. were collected andaftereach fraction the concentration of MeOH in the eluent was increased by 0.025 percent. A total of 16 fractions were collected. Fractions 3-9 were shown bytlc. to be pure. andwere combined to give 3.90 g. of a. glass consistingof 9. The sampel could not be crystallized. An" infraredshowed bands at 1750' cm. (oxime ester), 1730 cm. (lactone), and 1660 cm. (amide).

The reaction schemefor Examples VI'and VII is as follows:

Anal. calcd. for C H N O (964.257 C, 63.53; H, 8 9

N, 4. Found: C, 63.62; H, N 4

The reaction scheme for Examples Vlll and [X is as follows:

was added 4.00 g. (5.55 mmol.) of erythromycin B oxme. The mixture was stirred briefly to dissolve the COC].

3 2 Z L CH cn CH CI CH3 CH3 COCl H oxime and then was allowed to stand at 25 C. for four CH CH hours. The solution was then washed with 50 .ml. of 3 water containing 3 ml. of concentrated NH OH. The solvent was dried over anhyd. Na SO and evaporated 15 to give 5.24 g. of crude product.

Eboxlme 5 Purification of the crude product was accomplished by chromatography. A column of 140 g. of silica gel 3 3 was prepared in CH Cl and was made basic by eluting Cocl with 1.5 liter of CH Cl :0.3% triethylamine. The sam- 8 2O ple of crude 11 (5.24 g.) was placed on the column in 15 ml. of CH Cl :0.3%Et N and elution was begun with that solvent. Fractions of 125 ml. were collected and H after each fraction the concentration of MeOH in the CH3 eluent was increased by 0.025%. A total of 48 fractions c were collected. Fractions 16-28 were shown by tlc. to be pure and were combined to give 2.52 g. of 11. The sample could not be crystallized. An infrared analysis showed bands at 1750 cm. (oxime ester), 1730 cm. (lactone), and 1628 cm. (amide). For analysis the sample was concentrated twice by evaporation from ethanol solutions and then dried.

' Anal. calcd. for c u mo (978.240):

C, 62.62; H. 8.55; N, 4.30; 0 24.53. Found: C, 62.70; H, 8.80; N, 4.44; 0, 24.39.

40 EXAMPLE X [2,6-Dimethyl-4-(N-morpholinylcarbonyl)]benzoyl The reaction scheme for Examples X and XI is as fol- Chloride (l0) lows:

(\o COCl c 0 Q Et3N Eboxime CHgClz on H 3 3 CH cn COCl A CH Cl solution of the acid chloride 10 was pre- E pared in the following manner: To 100 ml. of CH Cl 0 was added 0.47] ml. (5.55 mmol.) of morpholine and \C/ 3.05 ml. (22.2 mmol.) of triethylamine. The mixture I was stirred and cooled to 0 C. in an ice-bath. Then, 1.00 ml. (5.55 mmol.) of 2,6-dimethylterephthaloyl chloride was added and after 30 minutes a sample was removed for infrared analysis which showed bands at CH3 CH3 1795 cm. (acid chloride) and at 1638 emf (amide). CH This solution was used directly without purification. t 3

EXAMPLE Xl CH 9-(0)-[2,6-Dimethyl-4-(N-morpholinylcarbo- H0 nyl)]benzoylerythromycin B Oxime (ll) fl Eb E 'To the CH Cl solution of 10 (Example X) at 0 C. -13- EXAMPLE XII A mixture of 50 ml. of dry CH Cl and 1.52 ml. (10.9 mmol.) of triethylamine was protected from moisture witha Drierite drying tube and was cooled to C. in an ice-bath. Then, 0.592 ml. (3.31 mmol.) of 2,6-dimethylterephthaloyl chloride was added, followed after a few minutes by 0.420 ml. (3.84 mmol.) of N-methylpiperazine added dropwise during three minutes. The ice-bath was removed and after 40 minutes 2.00 g. (2.73 mmol.) of erythromycin B oxime was added. After 24 hours, the mixture was diluted with 50 ml. CH Cl and was washed with two50 ml. portions of 1% NaHCO solution. The CH Cl was dried over Na SO and was evaporated to give 2.87 g. of crude product.

Before purification, a by-product of this reaction, the bis (erythromycin B oxime) ester of 2,6-dimethy1- terephthaloyl chloride was degraded to a mixture of erythromycin B oxime and 5. This was accomplished by allowing the crude product to stand at C. in 50 ml. of methanol, followed by 1 hour at reflux. The product was recovered from the methanol by evaporating, dissolving the residue in 150 ml. of CH C1 washing with 50 ml. of 1 percent NaHCO dryingover Na SO and finally evaporating the CH CI to leave 2.69 g. of crude product. A total of 8.0 g. of material was prepared in this way.

Thin layer chromatography showed the crude product contained six components. The desired product was separated using two column chromatography systems. The first system employed silica gel using CH Cl :0.9% Et N initially, followed with that eluent containing increasing (2,4,8 and 12 percent) amounts of acetonitrile. The fractions containing the major amount of the desired product (5.61 g.) were eluted with CH Cl :l2%CH CN:0.9%Et N. This material was rechromatographed in 2.8 g. portions on 200 g. of silica gel using EtOAc:15%MeOH: 1%Et N as the eluent. Fractions (12 ml.) were cut every minutes and were combined on the basis of tlc. results. Fractions of suitable purity were pooled and evaporated. The combined residue was dissolved in benzene, washed with 1 percent NaHCO and dried over Na SO Evaporation of the benzene gave 3.44 g. of 12 as a glass.

Anal. calcd. for C H N O (991.283):

EXAMPLE xnr 9-(0)-{2,6-Dimethyl-4-[N-(2-hydroxypropyl)-N- piperazinyl carbonyl]} benzoylerythromycin B Oxime A mixture of 100 ml. of CH CL and 1.09 ml. (6.00 mmol.) of 2,-dimethylterephthaloyl chloride was cooled to 0 C. in an ice-bath. In a ml. addition funnel was placed a mixture consisting of 3.04 ml. (21.8

mmol.) of triethylamine, 0.85 ml. (6.00 mmol.) of

was diluted with 50 ml. of'CH Cl washed with ml.

of 1 percent NaHCO solution, dried, and evaporated. After oil pumping, the residue weighed 5.75 g.

Purification of 4.0 g. of this'material was accomplished by chromatography of 2.0 g. samples on 200 g. columns of silica gel using EtOAc:l5%MeOH: l%Et N as the eluent. Fractions were combined on the basis of tlc. analysis. After evaporation of the eluent the residue was dissolved in ml. of benzene, which was then washed with 40 ml. of 1 percent NaHCO and 40 ml. of water. The benzene was dried over Na SO and evaporated to give 3.04 g. of 13 as a glass after dryin 24 hours in a vacuum oven at 65 C.

.01; H, 8.74; N, 5.65; O, 22.60. Found: C, 62.71; H, 8.71; N, 5.49; 0, 22.36. 50 Ana], calcd. f -1 1 0 103 5337 C, 62.65; H. 8.76; N, 5.41; 0, 23.18. The reaction scheme for Example X11 is as follows: Fwnd: 5'34; 2315' K\N/CH3 CH (\N/ 3 O\\C/N\/i COCl e Nd Et3N Eb-oxime CH CH H Z Z CH CH COCl COCl Eb-oxime coc1 EXAMPLE xiv 9-(0)-[2,6-Dimethyl-4-(4-methyl-l-pentyloxycarbonyl)]benzoylerythromycin B Oxime (14) A mixture of 100 ml. of CH Cl 1.52 ml. of (21.8

mmol.) of triethylamine, and 0.90 ml. (7.07 mmol.) of 4-methyl-l-pentanol was stirred at 25 C. while 1.18 ml. (7.07 mmol.) of 2,6-dimethylterephthaloyl chloride was added during a few minutes. After 2 hours at 25 C. a sample was examined and showed peaks at 1792 cm. (acid chloride) and 1718 cm. (ester) in the infrared. To this solution of 2,6-dimethy1-4-[4-methyl-lpentyloxycarbonyl] benzoyl chloride was added 4.00 g. (5.45 mmol.) of erythromycin B oxime. After stirring briefly the mixture was allowed to stand at 25 overnight. Then the mixture was washed with two 50 ml. portions of 1 percent NaI-ICO was dried over Na SO The solvent was evaporated to give 6.19 g. of crude product. I Two 2.0 g. samples of this product were chromatographed on 200 g. silica gel columns using benzenezSO percent ethylacetate:l%Et N as the eluent. Fractions of 8-10 ml. were cut every 8 minutes, and were pooled on the basis of tlc. results. After evaporation of the eluent, the residue was dissolved in benzene and wash washed with 1 percent NaHCO The benzene was dried over Na SO. and was evaporated to give 2.34 g. of 14 as a glass. The sampel was dried in a vacuum oven at 65 C. for 24 hours.

Anal. calcd. for C H N O (993.296)- c. 64.09; H. 8.93; N. 2.82; 0. 24.10. Found: C, 64.31: H, 9.13; N. 2.7610. 23.76.

The reaction scheme for Example XIV is as follows:

COCl

A/Y CH2C12 Et3N 3 I 3 COCl e Q Eb-Oxime E Q CH3 3 3 c 3 COCl 4 O 3 (EH3 EXAMPLE XV 9-(0)-{2,6-Dimethyl-4-[3-(N,N- dimethylamino)propyloxy carbonyl]} benzoylerythromycin B Oxime (15) A mixture of50 ml. OfCHgClz, 1.52 ml. (21.8 mmol.) of triethylamine, and 0.416 ml. (3.54 mmol.) of 3- dimethylamino-l-propanol was stirred at 25 C. while 0.592 ml. (3.26 mmol.) of 2,6-dimethylterephthaloyl chloride was added during 15 seconds. The mixture was stirred at 25 C. for 1 hour. A sample was removed which showed bands at 1792 cm. (acid chloride) and 1719 cm. (ester) in the infrared. To this solution of 2,6-dimethyl-4-[3-(N,N-dimethylamine)-propyloxycarbonyl]benzoyl chloride was added 2.00 g. (2.72 mmol.) of erythromycin B oxime. After 5 hours at 25 C. the mixture was diluted with 50 ml. of CH C| and was washed with 25ml. of 1 percent NaHCO The wash was separated and extracted with 30 ml. of CH Cl The combined CH Cl layers were dried over Na SO and evaporated to leave 3.21 g. of crude prodnet.

The sample was chromatographed in two portions on a 200 g. column of silica gel using EtOAc:l5%MeOH:- l%Et N as the eluent. Fractions of 8-10 ml. were collected every 8 minutes and were pooled on the basis of tlc. results. The eluent was evaporated and the residue was dissolved in ml. of benzene. The benzene was washed with 25 ml. of 1 percent NaHCO followed by 10 ml. of water. After drying over Na SO the benzene was evaporated to give 2.42 g. of 15 as a glass. The glass was dried in.a vacuum oven at 73 C. for 3 days.

3): Found: C, 62.98; H, 9

13 The reaction scheme for Example XV is as follows:

COC].

a no m 2 2 CH3 Et3N CH3 0o COCl CH CH3 1 3 l0 0 C, 7 Q CH3 CH EXAMPLE -XV1 9-(0)-{2,6-Dimethy1 4-[N-(4-methyl-l-penty1)carbamyl] benzoylerythromycin B Oxime (16) Following the procedure of Examples V111 and IX, but replacing n-butylamine with 1-amino4- methylpentane gave 16.

Anal. calcd. for C l-1 N 0 (992.312):

C, 64.15; H, 9.04; N, 4.23; 0. 22.57. Found: C, 64.00; H, 9.24; N, 4.17; 0, 22.36.

The reaction scheme for Example XVI is as follows: 40

COCl

CH3 CHgClz CH CH 0 3 COCl 3 o 1''! CH3 0 3 NM CH *0 3 Eb-oxime Q 3 3 011 c11 com CH 3. HO-.

EXAMPLE XVlI 9-(0)-{2,6-Dimethyl-4-[3-(N,N dimethylamino)propyl carbamyl]}benzoylerythromycin B Oxime (17) To a mixture of ml. 0fCH Cl and 3.82 ml. (27.3 mmol.) of triethylamine, cooled! in an ice-bath, was added 1.36 ml. (7.50 mmol.) of 2,6-dimethylterephthaloyl chloride. Immediately, 0.93 ml. (7.50 mmol.) of 3-dimethy1amino-l-propylamine was added. The icebath was removed and the mixture was stirred at 25 C. for 45 minutes. A sample showed bands at 1780 cm. (acid chloride) and 1655 cm." (amide) in the infrared. Then, to the solution of 2,6-dimethyl-4-[3 -(N,N-' dimethylamino)propylcarbamyl]henzoyl chloride was added 5.00 g. (6.82 mmol.) of erythromycin B oxime. The mixture was stirred at 25 C. for 5.5 hours. Then, washing with three 50 ml. portions of 1 percent Nal-l- CO drying over Na SO.,, and evaporating gave 7.04 g. of crude product.

The sample was chromatographed in two portions on a 200 g. silica gel column, using EtOAc:50%MeOl-l:- 1%Et N as the eluent. Fractions of 10-14 ml. were collected every 24 minutes, and were pooled on the basis of t1c..results. The eluent was evaporated to give 5.47 g. total of purified 17.

Anal. calcd. for C H N O (993.299):

C, 62.88; H. 8.93; N, 5.64; 0. 22.56. Found: C. 63.15; H, 9.17; N. 5.56;.0, 22.70.

The reaction scheme for Example XVll is as follows:

COCl

E 3 H NWN 2 c CHzClz CH3 CH3 Et3N COCl 0 iZ ii o 3 C \CH3 /N \CH3 52%,. Q 3 3 CH c11 Q (I) CH3 EXAMPLE XVlll 9-(0)-( 2,6-Dimethyl-4-carboxy )benzoylerythromycin B Oxime 18) A 1.00 g. of sample of 7 was dissolved in ml. of prehydrogenated ethanol. The mixture was stirred in the presence of 0.5 g. of 5% pd.-CaCO in an atmosphere of hydrogen. After 2hours the theoretical volume of hydrogen was consumed. The mixture was fi1- tered and the e thanol was evaporated to give 0.90 g. of

18 as a glass. The sample was dissolved in 18 ml. of

warm CH CI and 10 ml. of amyl acetate was added. On standing crystals formed giving 0.73 g. of pure 18 mp. l57-l 68 C.

EXAMPLE XIX Chlorination of Mesitoyl Chloride A 19.66 g. (0.108 mol.) sample of mesitoyl chloride was placed in a three neck flask fitted with a condenser,

gas inelt tube, and a magnetic stirring bar. The liquid was stirred and heated to 130-140 C. in an oil-bath. The flask was irradiated with a 150 watt flood lamp while chlorine gas was bubbled into the liquid. After 45' 'minutes the weight increase was 3.96 g. (3.74 g. theory for monochlorination). Further treatment with chlorine was stopped and the liquid was distilled through a 50 cm. Podbielniak column at 12.5 mm. Hg., bp. l47-149 C. to give 11.2 g. of product.

Anal. calcd. for CNH O CI (217.097):

4.64; CI., 32.66. Found: C, 55.07; 4.6

H. H, 8; CL, 32.52.

EXAMPLE XX 9-(0)-[2,6-Dimethyl-4-(N,N- dimethylaminomethyl)]benzoylerythromycin B Oxime 20 To a mixture of 200 ml. of Cl-l Cl 8.00 g. (10.9 mmol.) of erythromycin B oxime, and 6.1 ml. (43.6 mmol.) of triethylamine, stirred at C., was added 2.33 ml. (13.1 mmol.) of the mixture of 2 and 4- chloromethyl isomers of Example XIX. After standing for 5 hours at 25 C., the mixture was washed with two 70 ml. portions of 1 percent NaHCO was dried over Na SO and was evaporated to give 10 g. of crude isomeric oxime esters. This mixture was used without purification.

The sample of isomeric oxime esters was dissolved in a mixture of 500 ml. of methanol and 20 g. of dimethylamine. The solution was allowed to stand at 25 C. for

6 hours. Then, the methanol was concentrated to a volume of 50 ml. using a rotary evaporator and a water bath at 35 C. this residue was mixed with 250 ml. of

benzene and was washed with one 100 ml. and three 50 ml. portions of 1 percent NaI-ICO The combined washes were back extracted with 100 ml. of benzene. The combined benzene layers were dried over Na SO and were evaporated to give 8.16 g. of a mixture of 20 and erythromycin B oxime. The sample was dissolved in 25 m1. of CI-ICl and 70 ml. of hexane was added. On standing 3.72 g. of crystalline erythromycin B oxime was removed. The filtrate was evaporated to give 5.28 g. of a residue consisting of predominately 20. Further V purification was achieved by column chromatography of 2.6 g. samples of 200 g. columns of silica gel, using EtOAc: 10%MeOH :l%Et N as the eluent. Fractions of 6-8 ml., out every 8 minutes. were pooled on the basis of tlc. results. The eluent was evaporated and the residue was dissolved in ml. of benzene. The benzene was washed with three 50 ml. portions of NaHCO was dried over Na SO and was evaporated to give 1.67 g. of 20 free of erythromycin B oxime. Final purification was accomplished by removal of a small amount of 2,4- dimethyl-6-dimethylaminomethyl-N,N-dimethylbenzamide by chromatography on a 84 X 1.5 cm. column of Sephadex LI-I-20, using methanol as the eluent. The

.eluent was evaporated from pooled fractions and the residue was dissolved in'80 m1. of benzene. The benzene was washed with 30 ml. of dilute aqueous NaOH solution, was dried over N a SO and was evaporated to give 0.82 g. of pure 20.

Anal. calcd. for C ,,l-l N O (922.219):

C, 63.82; H, 9.07; N, 4.5 Found: C, 63.90; H, 9.31; N, 4.4

The reaction scheme for Examples XIX and XX is as follows:

+ Eb-oxime EX-AMPLE XXI 9-(0)-(2,4,6=Trimethyl)benzoylerythromycin A Oxime Hydrochloride (21) A 3.00 g. sample of 2 was dissolved in 50 ml. of methanol and 6.7 ml. of 0.5 N methanolic hydrochloric acid was added. The solventwas evaporated and the residue was dried for 2 hours in a vacuum oven at65 C.

Anal. calcd. for C H N o Cl (931.612):

CL, 3.92. Found: CL, 3.38.

EXAMPLE XXII 9-(0)-(2,4,6-Trimethyl)benzoylerythromycin A Oxime Acetic Acid Salt (22) A 3.00 g. sample of 2 was dissolved ina mixture of 40 ml. of benzene and 10 ml. of methanol. Then 0.195 ml. of acetic acid was added and the mixture was concentrated. The residue was dried with an oil pump. Examination of the sample by nmr. showed a three proton singlet at 2.01 ppm. consistent with the acetic acid salt.

Eb-oxime- COCl HN(Me) 2 MeOH EXAMPLE XXIII 9-(0 2 ,6-Dimethyl-4-(n-butyloxycarbonyl]} benzoylerythromycin' B Oxime(23) Following theprocedure of. Example XIV, but replacing 4-methyl-l-pentanol withn-butanol, gave 23.

Anal. calcd. for C H N O (965.241):

. C, 63.46; H, 8.77; N, 2 90; 0, 24 86 Found: C, 63.00; H, 8.83; N, 2 90; 0, 24 63 The compounds were tested for their activity against gram positive and gram negative bacteriain an agar dilution test. Results are reported in Tables Ithrough IV and are given in MIC values (minimum inhibitory concentrations) expressed'in micrograms ml. as to compounds from various Examples. Results in parentheses are those additional tests run which are log dilutions The pH of the agar dilution was 7.4.

The resulting of in vivo testing are reported in Table TABLE 11 rContinued IN VITRO BIOLOGICAL TEST RESULTS I' t-J Ery. A Ery. B Cpd. Cpd. 7 Cpd. 9 Cpd. 11 ltmagrt'llm rad/virus Static 100 100 100 100 100 100 Panugrelltts rt'divivus Cidal 100 100 100 100 100 100 'I'rt'chamtmas vagt'nalis C1M1 100 100 100 100 100 100 Crithidia fasciculata 100 100 100 100 100 100 Haenwphilus influenzae 9334 2.5 5.0 10.0 100 25 TABLE 111 IN VITRO BIOLOGICAL TEST RESULTS Bacteria Ery. A Ery. B Cpti. 2 Cpd. 4 Staph aureus 9144 0.2 0.39 0.78 0.39 Staph aureus Smith 0.2 0.39 0.39 0.39 Staph aureus Smith ER 100 100 100 100 Staph aureux Quinones 100 100 100 100 Staph aureus Wise 155 100 100 25 25 Strepfaecalis 10541 0.05 0.05 0.39 0.1 E. coli Juhl 50 100 100( 1000) 100(1000) Klebsiella pneumoniae 10031 6.2 3.1 50 25 Proteus vutgaris Abbott .U 100(1000) 100(1000) 100( 1000) 100(1000) Proteus mirabih's Finland No. 9 100(1000) 100(1000) 100( 1000) 100(1000) Salmonella typhimurium ED N0. 9 50 I00 100(1000) 50 Shigella sonnei 9290 25 25 100 25 Pseudomonas aerttgt'nosa BMH N0. 10 100 100 100 100 Strep pyogenes Roper 100 100 100 100 Strep pyagenes Scott 100 100 100 100 Additional Microorganisms Myco. gallt'se ticum S6 0.05 0.1 2.5 1.0 Myra. iranu arum 19168 0.02 0.02 0.25 0.25 Myco. yorhinis 17981 25 100 2.5 1.0 Myco. pneumoniae FH 0.01 0.02 0.02 0.02 Pan. redivivux Static 100 100 100 100 Pan. redivivus Cidal 100 100 100 100 Trichomonas vaginalt's C1M1 100 100 100 100 Crithidia fasiculata 100 100 100 100 Haemophilus influenzae 9334 2.5 5.0 25 25 TABLE [V IN VITRO BIOLOGICAL TEST RESULTS Bacteria Ery. A Ery. B Cpd. 12 Cpd. 18 Cpdv 14 Cpd. 17 Cpd. 20 Cpd. 13 Cpd. 1S Staph attrt'us 9144 0.39 0.39 1.56 12.5 12.5 6.2 0.39 6.2 3.1 Staph aureus Smith 0.39 0.39 1.56 12.5 12.5 3.1 0.39 6.2 3.1 Staph aureux Smith ER 100 100 100 100 100 100 100 100 Staph aureux Quinones 100 100 100 100 12.5 100 25 100 Staph aureus W156 155 100 100 100 12.5 100 12.5 100 25 Strep faec'alix E. t'nlt' 100 100 100 100 100 100 Juhl 50 100 (1000) 1000) 1000) 1000) 100 (10001 100(1) Klrlutrllu IIH'IIIHUII/tlf' 1011.11 (1.2 (1.2 100 101) 100 1110 12.5 100 50 Pmtattx vulgarix 100 100 100 100 100 100 100 100 100 Abbuttl] (1000) (1000) 1000) 1000) 1000) 1000) 1000) 1000) 1000) TABLE IV Continued IN VITRO BIOLOGICAL TEST RESULTS Bacteria Ery. A

Ery. B

Cpd. 12 Cpd. 18 Cpd. 14 Cpd. 17

Cpd. 20

Cpd. 13

Cpd. 15

Proteus mirabilix Finland No. 9

Salmonella Iyphimurium ED N0. 9

Shigella sonnei 9290 Pseudomonas aeruginosa BMH No. 10

Strep pyogenes Roper Smp pyogenex Scott Haemuphilus in fluenzae 93 34 Haenmphilux in flm'nzae Brimm CSF Brimm CS F Haemophilus influenzae Illinois Haemophilus in fluenzae Patterson Haemophilus influenzae She mwell Haemophilus in fluenzae Terry Additional Microo rgan- 1sms M ym. grunulamm Myco. I1 vor/1 1111.1

Trit/mmonus vagina/is C1 M1 CriI/zidia fascicw lulu 3 ,869 ,4414 a I 25 215 The erythromycin derivativeshere were also tested in vivo. They were orally administered to mice totreatan induced S taphylococcusaureus infection. Results are as follows:

TABLE V Approximate (.de sosamin e) Compound No. CD50 (Mg/Kg.)

l 100-200 3 50-100 5 100 clladinose) 7 200400 9 200-400 11 100-200 2 150-3.00 4 300 y l2 |50-300 l5 (erythronolide) l8 300 14 150-300 wherePR ishydrogenor hydnoxy, sandlR is a bulky alkyl lg 2 group of i4 carbons,,or a substituted .aryl ,group, said i3 300 substituted ar-ylgroup having -the structural formula 15 300 21 150 '20 22 200 As noted above prior art estersof erythromycin oximes are subject to unwanted hydrolysis. Such esters are quite reactive toward nucleophi'les such as water, alcohols and primary and secondary amines including the NH2-terminfll g p of amino fi flr n pcp- .where R andR are loweralkyl of 'l-4 oarbonsand lh tides. In many cases, such hydrolysis reaction-is facile i elected from the group consisting of carboxamide, and Occurs readily at room P a -C,C .alkyl, carhoxyalkyl, carboxy'lic acid, aminoal- With regard to the compounds of theinvention, it has ,ky], b tit t d i -lk j, j-b x 'i lkyl, a d been noted-that the rate of hydrolysis of: these esters is salts of flower. carboxylic acids. materially decreased. This is no doubt due to the in- 2,"-Th compound f l i 1 hi h i 9-(()) (2;6- crease of steric bulk about the ester carbonyl. dimethylJ-carbomethoxy)benzoylerythromycin B In order to demonstrate the difference in solvolysis of ,35 i prior art esters and the esters hereytypical esters were 3. The compound of ,claim 1 -which .is 9-(0) -(.2,'6- chosen as well as a known ester of erythromycin oxime .-dimethyl-'4-benzyloxycarbonyl)benzoylerythromycin"B and subjected to solvolysis by methanol The solvolysis oxime. reaction rates were followed by thin-layer chromatog- .4. The compound .of claim .1 which is-9-(0)-[.2.6- raphy, and the times required to half-solvolyse the esdimethyl 4-(l lm butylcarbomyl).1benzoylerythromycin ters were noted. Results are as follows: '8 oxime.

TABLE VI Comparison of Methanolysis Rates of Various Erythromycin Oxime sters 0 B:N-0-ll (Prior Art.) 1 day 9 /CH3 B:NOC-C;-CH (Example "'I)' 10 days CH3 CH3 B:N 0 ('l-- H3 (Example II) No reaction after 40 days CH lclaim: 5. The compound of claim .1 which is 9-(0)-.[2;6-

l. The compound 9-(0)(2,6-dimethyl-4- .dimethyl-4-(N-morpholinylcarbonyl)]benzoylerythrocarbomethoxy)benzoylerythromycin A oxime having wmycinB oxime. the following general structural formula 6. The compound of claim 1 which is Hm-(2.6-

zoylerythromycin B oxime. 11. The compound of claim 1 which is 9-(O)-{2,6- dimethyl-4-[3-(N,N-dimethylamino)propyl carbamyl]}benzoylerythromycin B oxime.

12. The compound of claim 1 which is 9-(0)-(2,6- dimethyl-4-carboxy)benzoylerythromycin B oxime.

13. The compound of claim 1 which is 9-(0)-[2,6- dimethyl-4-(N,N-dimethylaminomethyl)]benzoylerythromycin B oxime.

14. The compound of claim 1 which is 9-(0)-{2,6- dimethyl-4-[n-butyloxycarbonyl]}benzoylerythromycin B oxime. 

1. THE COMPOUND 9-(10)-(2,6-DIMETHYL-4CARBOMETHOXY)BENZOYLERYTHROMYCIN A OXIME HAVING THE FOLLOWERING GENERAL STRUCTURAL FORMULA
 2. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-carbomethoxy)benzoylerythromycin B oxime.
 3. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-benzyloxYcarbonyl)benzoylerythromycin B oxime.
 4. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(N-n-butylcarbomyl))benzoylerythromycin B oxime.
 5. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(N-morpholinylcarbonyl))benzoylerythromycin B oxime.
 6. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(N-methylpiperazinylcarbonyl)benzoylerythromycin B oxime.
 7. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(N-(2-hydroxypropyl)-N-piperazinyl carbonyl))benzoylerythromycin B oxime.
 8. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(4-methyl-1-pentyloxycarbonyl))benzoylerythromycin B oxime.
 9. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(3-(N, N-dimethylamino)propyloxy carbonyl))benzoylerythromycin B oxime.
 10. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(N-(4-methyl-1-pentyl)carbamyl))benzoylerythromycin B oxime.
 11. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(3-(N,N-dimethylamino)propyl carbamyl))benzoylerythromycin B oxime.
 12. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-carboxy)benzoylerythromycin B oxime.
 13. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(N,N-dimethylaminomethyl))benzoylerythromycin B oxime.
 14. The compound of claim 1 which is 9-(0)-(2,6-dimethyl-4-(n-butyloxycarbonyl))benzoylerythromycin B oxime. 